Ferroptosis is a recently recognized form of non‐apoptotic cell death caused by an iron‐dependent accumulation of lipid hydroperoxides, which plays important roles in a wide spectrum of pathological conditions. The present study was aimed to investigate the impact of ferroptosis on cisplatin‐induced sensory hair cell damage. Cell viability was determined by Cell Counting Kit‐8 and lactase dehydrogenase assays. The reactive oxygen species (ROS) levels were evaluated by 2,7‐Dichlorodi‐hydrofluorescein diacetate (DCFH‐DA) and MitoSox‐Red staining. Mitochondrial membrane potential (MMP) was measured by tetramethylrhodamine methyl ester (TMRM) staining. Lipid peroxidation, intracellular and mitochondrial iron were detected by Liperfluo, C11‐BODIPY^581/591, FerroOrange and Mito‐FerroGreen, respectively. We found that cisplatin treatment not only markedly augmented ROS accumulation, decreased the MMP, but increased lipid peroxidation and iron accumulation in House Ear Institute‐Organ of Corti 1 (HEI‐OC1) cells. Of note, treatment with the specific ferroptosis inhibitor ferrostatin‐1 could effectively abrogate the cisplatin‐induced toxicity and subsequent cell death. Specifically, the improvement of mitochondrial functions is important mechanisms for protective action of ferroptosis inhibitor against cisplatin‐induced damages in HEI‐OC1 cells. Moreover, inhibition of ferroptosis significantly protected murine cochlear hair cells against cisplatin damage. In addition, treatment murine cochlear hair cells with ferroptosis inducer, RSL3, significantly exacerbated cisplatin‐induced damage, which could be alleviated by ROS inhibitor N‐acetyl‐L‐cysteine. Collectively, our study indicated that ferroptosis inhibition could alleviate the cisplatin‐induced ototoxicity via inactivation of lipid peroxide radical and improvement of mitochondrial function in hair cells.

中文翻译：

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抑制铁锈病可以保护House Ear Institute-Corti 1细胞和耳蜗毛细胞器官免受顺铂诱导的耳毒性。

Ferroptosis是近来公认的一种非凋亡性细胞死亡形式，它是由铁依赖性脂质氢过氧化物的积累引起的，它在广泛的病理状况中起着重要的作用。本研究的目的是研究受铁症对顺铂诱导的感觉毛细胞损伤的影响。细胞活力由Cell Counting Kit-8和乳糖酶脱氢酶测定法确定。活性氧（ROS）水平通过2,7-二氯二氢荧光素二乙酸酯（DCFH-DA）和MitoSox-Red染色进行评估。线粒体膜电位（MMP）通过四甲基罗丹明甲酯（TMRM）染色测量。通过Liperfluo，C11-BODIPY ^581/591检测脂质过氧化，细胞内和线粒体铁，FerroOrange和Mito-FerroGreen。我们发现顺铂治疗不仅显着增加了房耳研究所的Corti 1器官（HEI-OC1）细胞中的ROS积累，降低了MMP，而且还增加了脂质过氧化作用和铁积累。值得一提的是，使用特定的ferroptosis抑制剂ferrostatin-1治疗可以有效消除顺铂引起的毒性和随后的细胞死亡。具体而言，线粒体功能的改善是铁蛋白抑制剂在对抗HEI-OC1细胞中顺铂诱导的损伤中的保护作用的重要机制。此外，抑制铁锈病显着保护了鼠耳蜗毛细胞免受顺铂损害。此外，用肥大病诱导剂RSL3处理鼠耳蜗毛细胞会大大加重顺铂诱导的损伤，ROS抑制剂N-乙酰-L-半胱氨酸可以缓解这种情况。总体而言，我们的研究表明，抑制铁锈病可以通过使脂质过氧化物自由基失活和改善毛细胞中的线粒体功能来减轻顺铂引起的耳毒性。